Abstract
Cerebellar granule neurons (CGN) cultured in the presence of serum and depolarizing potassium concentrations undergo apoptosis when switched to serum-free medium containing physiological potassium concentrations. Here we show that processing of the key protease, caspase-3, depends on the activation of caspase-9, but not of caspase-8. Selective peptide inhibitors of caspase-9 block processing of caspase-3 and caspase-8 and inhibit apoptosis, whereas a selective inhibitor of caspase-8 blocks neither processing of caspase-3 nor cell death. The data obtained with peptide inhibitors were confirmed by adenovirally mediated ectopic expression of the cytokine response modifier A (crmA), the baculovirus protein p35, and the X chromosome-linked inhibitor of apoptosis (XIAP). Further, caspase-8-activating death receptors do not mediate apoptosis in CGN and potassium withdrawal-induced apoptosis evolves unaltered in gld or lpr mice, which harbor mutations in the CD95/CD95 ligand system. Thus, neuronal apoptosis triggered by potassium deprivation is death receptor-independent but involves the mitochondrial pathway of caspase activation.
Copyright 2001 Academic Press.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Animals
-
Apoptosis / drug effects
-
Apoptosis / physiology*
-
Caspase 3
-
Caspase 8
-
Caspase 9
-
Caspase Inhibitors
-
Caspases / metabolism*
-
Cell Survival / physiology
-
Cells, Cultured
-
Cerebellum / cytology*
-
Coumarins / pharmacology
-
Enzyme Inhibitors / pharmacology
-
Enzyme Precursors / antagonists & inhibitors
-
Enzyme Precursors / metabolism
-
Gene Expression Regulation, Viral
-
Inhibitor of Apoptosis Proteins
-
Mice
-
Mice, Inbred BALB C
-
Mice, Inbred C57BL
-
Mice, Inbred MRL lpr
-
Neurons / cytology*
-
Neurons / enzymology*
-
Oligopeptides / pharmacology
-
Potassium / pharmacology*
-
Proteins / genetics
-
Rats
-
Rats, Sprague-Dawley
-
Serpins / genetics
-
Specific Pathogen-Free Organisms
-
Transgenes / physiology
-
Viral Proteins / genetics
-
X-Linked Inhibitor of Apoptosis Protein
Substances
-
Caspase Inhibitors
-
Coumarins
-
Enzyme Inhibitors
-
Enzyme Precursors
-
Inhibitor of Apoptosis Proteins
-
Oligopeptides
-
Proteins
-
Serpins
-
Viral Proteins
-
X-Linked Inhibitor of Apoptosis Protein
-
acetyl-aspartyl-glutamyl-valyl-aspartyl-amino-4-methylcoumarin
-
inhibitor of apoptosis, Nucleopolyhedrovirus
-
interleukin-1beta-converting enzyme inhibitor
-
Casp3 protein, mouse
-
Casp3 protein, rat
-
Casp8 protein, mouse
-
Casp8 protein, rat
-
Casp9 protein, mouse
-
Casp9 protein, rat
-
Caspase 3
-
Caspase 8
-
Caspase 9
-
Caspases
-
Potassium